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The S-palmitoylome ...
The S-palmitoylome and DHHC-PAT interactome of Drosophila melanogaster S2R+cells indicate a high degree of conservation to mammalian palmitoylomes
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- Porcellato, Elena (författare)
- Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.
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- Gonzalez-Sanchez, Juan Carlos (författare)
- Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.;Heidelberg Univ, BioQuant, Heidelberg, Germany.
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- Ahlmann-Eltze, Constantin (författare)
- Univ Heidelberg ZMBH, Ctr Mol Biol, Heidelberg, Germany.;EMBL, Genome Biol Unit, Heidelberg, Germany.
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- Elsakka, Mahmoud Ali (författare)
- Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.
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- Shapira, Itamar (författare)
- Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.
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- Fritsch, Juergen (författare)
- Univ Klinikum Schleswig Holstein, Inst Immunol, Kiel, Germany.;Univ Hosp Regensburg, Dept Infect Prevent & Infect Dis, Regensburg, Germany.
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- Navarro, Juan Antonio (författare)
- Univ Regensburg, Inst Zool, Regensburg, Germany.;INCLIVA Biomedial Res Inst, Valencia, Spain.;Univ Valencia, Dept Genet, Valencia, Spain.
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- Anders, Simon (författare)
- Univ Heidelberg ZMBH, Ctr Mol Biol, Heidelberg, Germany.
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- Russell, Robert B. (författare)
- Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.;Heidelberg Univ, BioQuant, Heidelberg, Germany.
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- Wieland, Felix T. (författare)
- Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.
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- Metzendorf, Christoph (författare)
- Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.
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Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany Heidelberg Univ, Biochem Ctr BZH, Heidelberg, Germany.;Heidelberg Univ, BioQuant, Heidelberg, Germany. (creator_code:org_t)
- 2022-08-12
- 2022
- Engelska.
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 17:8
- Relaterad länk:
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https://doi.org/10.1...
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https://uu.diva-port... (primary) (Raw object)
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Protein S-palmitoylation, the addition of a long-chain fatty acid to target proteins, is among the most frequent reversible protein modifications in Metazoa, affecting subcellular protein localization, trafficking and protein-protein interactions. S-palmitoylated proteins are abundant in the neuronal system and are associated with neuronal diseases and cancer. Despite the importance of this post-translational modification, it has not been thoroughly studied in the model organism Drosophila melanogaster. Here we present the palmitoylome of Drosophila S2R+ cells, comprising 198 proteins, an estimated 3.5% of expressed genes in these cells. Comparison of orthologs between mammals and Drosophila suggests that S-palmitoylated proteins are more conserved between these distant phyla than non-S-palmitoylated proteins. To identify putative client proteins and interaction partners of the DHHC family of protein acyl-transferases (PATs) we established DHHC-BioID, a proximity biotinylation-based method. In S2R+ cells, ectopic expression of the DHHC-PAT dHip14-BioID in combination with Snap24 or an interaction-deficient Snap24-mutant as a negative control, resulted in biotinylation of Snap24 but not the Snap24-mutant. DHHC-BioID in S2R+ cells using 10 different DHHC-PATs as bait identified 520 putative DHHC-PAT interaction partners of which 48 were S-palmitoylated and are therefore putative DHHC-PAT client proteins. Comparison of putative client protein/DHHC-PAT combinations indicates that CG8314, CG5196, CG5880 and Patsas have a preference for transmembrane proteins, while S-palmitoylated proteins with the Hip14-interaction motif are most enriched by DHHC-BioID variants of approximated and dHip14. Finally, we show that BioID is active in larval and adult Drosophila and that dHip14-BioID rescues dHip14 mutant flies, indicating that DHHC-BioID is non-toxic. In summary we provide the first systematic analysis of a Drosophila palmitoylome. We show that DHHC-BioID is sensitive and specific enough to identify DHHC-PAT client proteins and provide DHHC-PAT assignment for ca. 25% of the S2R+ cell palmitoylome, providing a valuable resource. In addition, we establish DHHC-BioID as a useful concept for the identification of tissue-specific DHHC-PAT interactomes in Drosophila.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
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Porcellato, Elen ...
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Gonzalez-Sanchez ...
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Ahlmann-Eltze, C ...
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Elsakka, Mahmoud ...
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Shapira, Itamar
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Fritsch, Juergen
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Navarro, Juan An ...
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Anders, Simon
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Russell, Robert ...
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Wieland, Felix T ...
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Metzendorf, Chri ...
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